WO2014094462A1 - Biological preparation method of (s)-3-methylamino-1-(2-thienyl)-1-propyl alcohol - Google Patents

Biological preparation method of (s)-3-methylamino-1-(2-thienyl)-1-propyl alcohol Download PDF

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WO2014094462A1
WO2014094462A1 PCT/CN2013/083235 CN2013083235W WO2014094462A1 WO 2014094462 A1 WO2014094462 A1 WO 2014094462A1 CN 2013083235 W CN2013083235 W CN 2013083235W WO 2014094462 A1 WO2014094462 A1 WO 2014094462A1
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thienyl
methylamino
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乐庸堂
鞠鑫
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苏州汉酶生物技术有限公司
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    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms

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  • the invention belongs to the field of biopharmaceuticals and green chemistry, and particularly relates to a biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol.
  • Duloxetine is a low side effect drug that can effectively treat mental disorders and metabolic disorders (US patent 5,023,269)
  • the key to the synthesis of duloxetine is to obtain the intermediate (S)-3-methylamino-1-(2-thienyl)-1-propanol (MMAA) containing a chiral center, thus asymmetric reduction 3-methylamino-1-( 2-Thienyl)-1-propanone (MMAK) is one of the most effective and researched methods available.
  • MMAA 3-methylamino-1-( 2-Thienyl)-1-propanone
  • the technical problem to be solved by the present invention is to provide an improved biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol.
  • the present invention adopts the following technical scheme: a biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol, which is 3-methylamino group 1-(2-Thienyl)-1-propanone is a substrate which undergoes asymmetric reduction in the presence of a biocatalyst, cofactor and cofactor regeneration system to form (S)-3-methylamino group- 1-(2-Thienyl)-1-propanol.
  • the biocatalyst is a ketoreductase
  • the cofactor regeneration system comprises glucose and glucose dehydrogenase
  • the asymmetric reduction reaction is carried out in an aqueous phase buffer having a pH of 6-8, in an initial state of reaction.
  • concentration of the substrate 3-methylamino-1-(2-thienyl)-1-propanone is 100-150 Mg/mL.
  • the mass ratio of the ketoreductase to the substrate 3-methylamino-1-(2-thienyl)-1-propanone is 1-2%.
  • the mass ratio of the cofactor: glucose:glucose dehydrogenase: 3-methylamino-1-(2-thienyl)-1-propanone is (0.0005-0.002). ): (1.2-2): (0.01-0.05): 1.
  • the cofactor is NAD/NADH or NADP/NADPH, and is preferably NADP/NADPH.
  • aqueous phase buffer is a triethanolamine buffer solution.
  • asymmetric reduction reaction was carried out in an aqueous phase buffer having a pH of 7.0.
  • the pH of the aqueous phase buffer in which the asymmetric reduction reaction is carried out is adjusted with a sodium hydroxide solution, a sodium carbonate solution or a sodium hydrogencarbonate solution.
  • the above preparation method can be carried out as follows: in the reaction vessel, a substrate 3-methylamino-1-(2-thienyl)-1-propanone, glucose, an aqueous phase buffer solution is sequentially added, and the mixture is uniformly stirred. The ketone reductase, glucose dehydrogenase and the cofactor are further added, and the reaction is stirred at 25-45 ° C, and the progress of the reaction is detected by HPLC. When the conversion rate reaches 90-99%, the reaction is terminated.
  • ketoreductase (Suzhou Hanase Biotechnology Co., Ltd., KRED137) and glucose dehydrogenase (Suzhou Hanzyme Biotechnology Co., Ltd., GDH102) in the present invention are commercially available.
  • the invention optimizes the process conditions, increases the substrate concentration, reduces the enzyme dosage, thereby reducing the production cost, realizing a high-efficiency biotransformation process, and is suitable for industrial production.
  • reaction formula of the present invention is as follows:
  • the reaction of NADP in triethanolamine buffer begins.
  • the reaction temperature was maintained at 30 °C.
  • the pH of the reaction solution was adjusted by dropwise addition of a 2 M NaOH solution through a pH titrator.
  • the pH is controlled at 7.0 and the initial control point is 6. 5, the final control point is 6.99.
  • the reaction was carried out for 23 h with a conversion of >99%.
  • the pH of the reaction solution was adjusted by dropwise addition of a 2 M NaOH solution through a pH titrator.
  • the pH is controlled at 7.0 and the initial control point is 6. 5, the final control point is 6.99.
  • the reaction was carried out for 23 h with a conversion of >98%.

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Abstract

The present invention provides a biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propyl alcohol, wherein 3-methylamino-1-(2-thienyl)-1-acetone is used as a substrate; under a condition that a biocatalyst ketoreductase, a cofactor and a cofactor regeneration system (comprising glucose and glucose dehydrogenase) exist, the substrate undergoes an asymmetric reduction reaction to generate the (S)-3-methylamino-1-(2-thienyl)-1-propyl alcohol; the asymmetric reduction reaction is carried out in a water-phase buffer solution with a pH of 6-8; and in a initial reaction system, concentration of the substrate is 100-150 mg/mL, and a mass ratio of the ketoreductase to the substrate is 1-2%. The present invention improves the concentration of the substrate and reduces enzyme usage.

Description

(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法  Biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol
技术领域：Technical field:
本发明属于生物制药和绿色化学领域，具体涉及一种(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法。The invention belongs to the field of biopharmaceuticals and green chemistry, and particularly relates to a biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol.
背景技术：Background technique:
度洛西汀(Duloxetine)是一种低副作用的，能够有效治疗精神紊乱和代谢紊乱的药物(US patent 5,023,269)，合成度洛西汀的关键是获得含有手性中心的中间体(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇(MMAA)，因此不对称还原3-甲胺基-1-( 2-噻吩基)-1-丙酮(MMAK)得到该中间体是目前最有效、研究最多的方法之一。在实现该途径的化学还原法中，由于需要钌金属催化剂催化下，加入4.5 MPa的氢气反应，其经济性、安全性和环境友好性并不能满足生产的需要(Tetrahedron Lett. 1990, 31:7101–7104)。利用扁桃酸作为拆分试剂的化学拆分法，由于得率低和经济性差等原因，也不能作为生产MMAA的主要生产方法(Tetrahedron Asymm. 2003, 14:1631–1636)。因此，利用生物酶法还原MMAK得到MMAA，以其高效率和环境友好性就成为目前最有效的方法。Duloxetine is a low side effect drug that can effectively treat mental disorders and metabolic disorders (US patent 5,023,269), the key to the synthesis of duloxetine is to obtain the intermediate (S)-3-methylamino-1-(2-thienyl)-1-propanol (MMAA) containing a chiral center, thus asymmetric reduction 3-methylamino-1-( 2-Thienyl)-1-propanone (MMAK) is one of the most effective and researched methods available. In the chemical reduction method to achieve this route, due to the need for ruthenium metal catalyst catalysis, add 4.5 The economical, safety and environmental friendliness of the MPa hydrogen reaction does not meet the production needs (Tetrahedron Lett. 1990, 31:7101–7104). The chemical resolution method using mandelic acid as a resolving agent cannot be used as the main production method for producing MMAA due to low yield and poor economy (Tetrahedron) Asymm. 2003, 14:1631–1636). Therefore, the use of bioenzymatic reduction of MMAK to obtain MMAA has become the most effective method with its high efficiency and environmental friendliness.
在生物还原法中，有利用Candida magnoliae重组还原酶催化该还原反应的实例，但是该方法催化剂加入量高，底物浓度过低，明显难以产业化(US patent 2008/0220484 A1)。In the biological reduction method, there is the use of Candida The magnoliae recombinant reductase catalyzes an example of the reduction reaction, but the method has a high catalyst addition amount and a low substrate concentration, which is obviously difficult to industrialize (US patent 2008/0220484 A1).
发明内容：Summary of the invention:
本发明所要解决的技术问题是提供一种改进的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法。The technical problem to be solved by the present invention is to provide an improved biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol.
为解决以上技术问题，本发明采用如下技术方案：一种(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其以3-甲胺基-1-(2-噻吩基)-1-丙酮为底物，该底物在生物催化剂、辅因子和辅因子再生***的存在下发生不对称还原反应生成(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇。生物催化剂为酮还原酶，所述的辅因子再生***包括葡萄糖和葡萄糖脱氢酶，所述的不对称还原反应在pH值为6-8的水相缓冲液中进行，在起始状态的反应体系中。底物3-甲胺基-1-(2-噻吩基)-1-丙酮的浓度为100-150 mg/mL。酮还原酶与底物3-甲胺基-1-(2-噻吩基)-1-丙酮的质量比为1-2%。In order to solve the above technical problems, the present invention adopts the following technical scheme: a biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol, which is 3-methylamino group 1-(2-Thienyl)-1-propanone is a substrate which undergoes asymmetric reduction in the presence of a biocatalyst, cofactor and cofactor regeneration system to form (S)-3-methylamino group- 1-(2-Thienyl)-1-propanol. The biocatalyst is a ketoreductase, the cofactor regeneration system comprises glucose and glucose dehydrogenase, and the asymmetric reduction reaction is carried out in an aqueous phase buffer having a pH of 6-8, in an initial state of reaction. In the system. The concentration of the substrate 3-methylamino-1-(2-thienyl)-1-propanone is 100-150 Mg/mL. The mass ratio of the ketoreductase to the substrate 3-methylamino-1-(2-thienyl)-1-propanone is 1-2%.
进一步地，在起始状态的反应体系中，辅因子：葡萄糖：葡糖糖脱氢酶：3-甲胺基-1-(2-噻吩基)-1-丙酮的质量比为（0.0005-0.002）：（1.2-2）：（0.01-0.05）：1。Further, in the initial reaction system, the mass ratio of the cofactor: glucose:glucose dehydrogenase: 3-methylamino-1-(2-thienyl)-1-propanone is (0.0005-0.002). ): (1.2-2): (0.01-0.05): 1.
进一步地，辅因子为NAD/NADH或NADP/NADPH，且优选为NADP/NADPH。Further, the cofactor is NAD/NADH or NADP/NADPH, and is preferably NADP/NADPH.
进一步地，水相缓冲液为三乙醇胺缓冲溶液。Further, the aqueous phase buffer is a triethanolamine buffer solution.
进一步地，不对称还原反应在pH值为7.0的水相缓冲液中进行。Further, the asymmetric reduction reaction was carried out in an aqueous phase buffer having a pH of 7.0.
进一步地，不对称还原反应所处水相缓冲液的pH值用氢氧化钠溶液、碳酸钠溶液或碳酸氢钠溶液调节。Further, the pH of the aqueous phase buffer in which the asymmetric reduction reaction is carried out is adjusted with a sodium hydroxide solution, a sodium carbonate solution or a sodium hydrogencarbonate solution.
进一步地，上述制备方法的实施过程可按照如下：在反应容器中依次加入底物3-甲胺基-1-(2-噻吩基)-1-丙酮、葡萄糖、水相缓冲液，搅拌均匀，继续加入酮还原酶、葡萄糖脱氢酶和所述的辅因子，在25-45℃下，搅拌反应，利用HPLC检测反应进程，待转化率达90-99%时，结束反应。Further, the above preparation method can be carried out as follows: in the reaction vessel, a substrate 3-methylamino-1-(2-thienyl)-1-propanone, glucose, an aqueous phase buffer solution is sequentially added, and the mixture is uniformly stirred. The ketone reductase, glucose dehydrogenase and the cofactor are further added, and the reaction is stirred at 25-45 ° C, and the progress of the reaction is detected by HPLC. When the conversion rate reaches 90-99%, the reaction is terminated.
本发明中的酮还原酶（苏州汉酶生物技术有限公司，KRED137）、葡萄糖脱氢酶（苏州汉酶生物技术有限公司，GDH102）均可商购获得。The ketoreductase (Suzhou Hanase Biotechnology Co., Ltd., KRED137) and glucose dehydrogenase (Suzhou Hanzyme Biotechnology Co., Ltd., GDH102) in the present invention are commercially available.
本发明的有益效果在于:The beneficial effects of the invention are:
本发明通过对工艺条件的优化，提高底物浓度，降低酶用量，从而降低了生产成本，实现了高效率的生物转化过程，适合产业化生产。The invention optimizes the process conditions, increases the substrate concentration, reduces the enzyme dosage, thereby reducing the production cost, realizing a high-efficiency biotransformation process, and is suitable for industrial production.
具体实施方式：detailed description:
本发明的反应式如下： The reaction formula of the present invention is as follows:
Figure PCTCN2013083235-appb-I000001
Figure PCTCN2013083235-appb-I000001
下面的实施例可以使本专业的技术人员更全面地理解本发明，但不以任何方式限制本发明。 The following examples are provided to enable those skilled in the art to understand the invention more fully, but are not intended to limit the invention in any way.
实施例一 Embodiment 1
向50 mL反应三口瓶中添加1.0 g底物MMAK和1.2g葡萄糖，再向反应瓶中加入8 mL配好的pH为7.0，0.1 M 三乙醇胺缓冲液。置于30℃，900 r/min磁力搅拌水浴锅中搅拌，并用2 M NaOH溶液调节pH至7.0。待温度稳定，向反应瓶中加入0.95 mL 溶有20 mg 酮还原酶，31 mg GDH的三乙醇胺缓冲液。用NaOH调节pH至7.0，并用0.95 mL三乙醇胺缓冲液冲洗并加入反应瓶中。再向反应瓶中加入100 μL溶有0.5 mg NADP的三乙醇胺缓冲液，反应开始。反应温度保持在30℃。反应液的pH通过pH滴定仪滴加2 M NaOH 溶液调节。控制pH在7.0，初控点为6. 5，终控点为6.99。定时取样供HPLC检测中控。反应23 h，转化率>99%。Add 1.0 g of substrate MMAK and 1.2 g of glucose to a 50 mL reaction three-necked flask, and add 8 to the reaction flask. The pH is 7.0, 0.1 M triethanolamine buffer. Stir at 30 ° C, 900 r / min magnetic stirring water bath, and use 2 M The pH of the NaOH solution was adjusted to 7.0. To stabilize the temperature, add 0.95 mL to the reaction flask and dissolve 20 mg of ketoreductase, 31 mg. GDH triethanolamine buffer. The pH was adjusted to 7.0 with NaOH and flushed with 0.95 mL of triethanolamine buffer and added to the reaction flask. Add 100 μL of 0.5 mg to the reaction flask. The reaction of NADP in triethanolamine buffer begins. The reaction temperature was maintained at 30 °C. The pH of the reaction solution was adjusted by dropwise addition of a 2 M NaOH solution through a pH titrator. The pH is controlled at 7.0 and the initial control point is 6. 5, the final control point is 6.99. Timed sampling for HPLC detection. The reaction was carried out for 23 h with a conversion of >99%.
实施例二Embodiment 2
向反应器中添加100.0 g底物MMAK和120.0 g葡萄糖，再向反应瓶中加入800 mL配好的pH为7.0，0.1 M 三乙醇胺缓冲液。置于30℃，机械搅拌水浴锅中搅拌，并用2 M NaOH溶液调节pH至7.0。待温度稳定，向反应瓶中加入95 mL 溶有2 g 酮还原酶，3.1 g GDH的三乙醇胺缓冲液。用NaOH调节pH至7.0，并用95 mL三乙醇胺缓冲液冲洗并加入反应瓶中。再向反应瓶中加入10 mL溶有50 mg NADP的三乙醇胺缓冲液，反应开始。反应温度保持在30℃。反应液的pH通过pH滴定仪滴加2 M NaOH 溶液调节。控制pH在7.0，初控点为6. 5，终控点为6.99。定时取样供HPLC检测中控。反应23 h，转化率>98%。Add 100.0 g of substrate MMAK and 120.0 g of glucose to the reactor and add 800 to the reaction flask. The pH is 7.0, 0.1 M triethanolamine buffer. Stir at 30 ° C in a mechanically stirred water bath and adjust the pH to 7.0 with 2 M NaOH solution. Wait for the temperature to stabilize, add 95 to the reaction flask mL was dissolved in 2 g ketoreductase, 3.1 g GDH in triethanolamine buffer. Adjust the pH to 7.0 with NaOH and use 95 Rinse with triethanolamine buffer and add to the reaction flask. Add 10 mL to the reaction flask and dissolve 50 mg. The reaction of NADP in triethanolamine buffer begins. The reaction temperature was maintained at 30 °C. The pH of the reaction solution was adjusted by dropwise addition of a 2 M NaOH solution through a pH titrator. The pH is controlled at 7.0 and the initial control point is 6. 5, the final control point is 6.99. Timed sampling for HPLC detection. The reaction was carried out for 23 h with a conversion of >98%.
上述实施例只为说明本发明的技术构思及特点，其目的在于让熟悉此项技术的人士能够了解本发明的内容并据以实施，并不能以此限制本发明的保护范围。凡根据本发明精神实质所作的等效变化或修饰，都应涵盖在本发明的保护范围之内。The above embodiments are merely illustrative of the technical concept and the features of the present invention, and the purpose of the present invention is to enable those skilled in the art to understand the present invention and to implement the present invention, and the scope of the present invention is not limited thereto. Equivalent variations or modifications made in accordance with the spirit of the invention are intended to be included within the scope of the invention.

Claims (7)

1、一种(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其以3-甲胺基-1-(2-噻吩基)-1-丙酮为底物，该底物在生物催化剂、辅因子和辅因子再生***的存在下发生不对称还原反应生成(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇，其特征在于：所述的生物催化剂为酮还原酶，所述的辅因子再生***包括葡萄糖和葡萄糖脱氢酶，所述的不对称还原反应在pH值为6-8的水相缓冲液中进行，在起始状态的反应体系中，所述的底物3-甲胺基-1-(2-噻吩基)-1-丙酮的浓度为100-150 mg/mL，所述的酮还原酶与所述的底物3-甲胺基-1-(2-噻吩基)-1-丙酮的质量比为1-2%。 A biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol, which is 3-methylamino-1-(2-thienyl)-1 - Acetone is a substrate which undergoes asymmetric reduction in the presence of a biocatalyst, cofactor and cofactor regeneration system to form (S)-3-methylamino-1-(2-thienyl)-1- Propyl alcohol, characterized in that the biocatalyst is a ketone reductase, the cofactor regeneration system comprises glucose and glucose dehydrogenase, and the asymmetric reduction reaction is buffered in an aqueous phase having a pH of 6-8. In the initial reaction system, the concentration of the substrate 3-methylamino-1-(2-thienyl)-1-propanone is 100-150. The mass ratio of the ketoreductase to the substrate 3-methylamino-1-(2-thienyl)-1-propanone is 1-2%.
2、根据权利要求1所述的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其特征在于：在起始状态的反应体系中，辅因子：葡萄糖：葡糖糖脱氢酶：3-甲胺基-1-(2-噻吩基)-1-丙酮的质量比为（0.0005-0.002）：（1.2-2）：（0.01-0.05）：1。The method for biologically preparing (S)-3-methylamino-1-(2-thienyl)-1-propanol according to claim 1, wherein in the initial reaction system, Cofactor: glucose: glucose sugar dehydrogenase: mass ratio of 3-methylamino-1-(2-thienyl)-1-propanone (0.0005-0.002): (1.2-2): (0.01-0.05) ):1.
3、根据权利要求1所述的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其特征在于：所述的辅因子为NAD/NADH或NADP/NADPH。The biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol according to claim 1, wherein the cofactor is NAD/NADH. Or NADP/NADPH.
4、根据权利要求1所述的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其特征在于：所述的水相缓冲液为三乙醇胺缓冲溶液。The method for biologically preparing (S)-3-methylamino-1-(2-thienyl)-1-propanol according to claim 1, wherein the aqueous buffer is three Ethanolamine buffer solution.
5、根据权利要求1所述的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其特征在于：不对称还原反应在pH值为7.0的水相缓冲液中进行。The method for biologically producing (S)-3-methylamino-1-(2-thienyl)-1-propanol according to claim 1, wherein the asymmetric reduction reaction is at a pH of 7.0. The aqueous phase buffer is carried out.
6、根据权利要求1、4及5中任一项所述的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其特征在于：不对称还原反应所处的水相缓冲液的pH值用氢氧化钠溶液、碳酸钠溶液或碳酸氢钠溶液调节。The method for biologically producing (S)-3-methylamino-1-(2-thienyl)-1-propanol according to any one of claims 1, 4 and 5, which is characterized in that The pH of the aqueous phase buffer in which the symmetric reduction reaction is carried out is adjusted with a sodium hydroxide solution, a sodium carbonate solution or a sodium hydrogencarbonate solution.
7、根据权利要求1或2所述的(S)-3-甲胺基-1-(2-噻吩基)-1-丙醇的生物制备方法，其中，所述的制备方法的实施过程如下：在反应容器中依次加入底物3-甲胺基-1-(2-噻吩基)-1-丙酮、葡萄糖、水相缓冲液，搅拌均匀，继续加入酮还原酶、葡萄糖脱氢酶和所述的辅因子，在25-45℃下，搅拌反应，利用HPLC检测反应进程，待转化率达90-99%时，结束反应。The biological preparation method of (S)-3-methylamino-1-(2-thienyl)-1-propanol according to claim 1 or 2, wherein the preparation process is as follows : Add the substrate 3-methylamino-1-(2-thienyl)-1-propanone, glucose, and aqueous buffer to the reaction vessel in sequence, stir evenly, and continue to add ketone reductase, glucose dehydrogenase and The cofactor described above was stirred at 25-45 ° C, and the progress of the reaction was measured by HPLC. When the conversion rate reached 90-99%, the reaction was terminated.
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Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103173503A (en) * 2013-04-10 2013-06-26 江西师范大学 Method for biologically preparing (S)-4-chloro-3-hydroxy butyric acid ethyl ester with recombinant escherichia coli expressed ketoreductase
CN103421854A (en) * 2013-07-30 2013-12-04 苏州汉酶生物技术有限公司 Biological preparation method of (S)-3-(dimethylamino)-1-(thiophene-2-radical)-1-propyl alcohol
CN103468757A (en) * 2013-09-18 2013-12-25 苏州汉酶生物技术有限公司 Biological preparation method of intermediate of atazanavir
CN106011096B (en) * 2016-07-27 2020-11-03 苏州汉酶生物技术有限公司 Engineered ketoreductase polypeptide and method for preparing (S) -3- (dimethylamino) -1- (thiophene-2-yl) -1-propanol by using same
CN107417663B (en) * 2017-09-25 2019-05-28 台州职业技术学院 A kind of preparation method of high-purity (S) -3- methylamino -1- (thiophene -2- base) propyl alcohol
CN110129307B (en) * 2019-05-23 2023-09-05 安徽趣酶生物科技有限公司 Immobilized ketoreductase mutant and application thereof in preparation of duloxetine chiral alcohol intermediate and analogue thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080220484A1 (en) * 2005-09-19 2008-09-11 Basf Aktiengesellschaft Novel Dehydrogenases, the Derivatives thereof, and Method for the Production of Optically Active Alkanols
CN101613672A (en) * 2009-08-04 2009-12-30 南京工业大学 A kind of recombination bacillus coli and construction process thereof of asymmetric conversion preparation (S)-4-chloro-ethyl 3-hydroxybutanoate
CN101993828A (en) * 2010-11-08 2011-03-30 江南大学 Method for improving transformation efficiency of (S)-phenyl glycol by coupling glucose-6-phosphate dehydrogenase and (S)-carbonyl reductase
CN102206686A (en) * 2011-04-19 2011-10-05 华东理工大学 Preparation method of methyl (R)-o-chloromandelate utilizing biocatalytic asymmetric reduction

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080220484A1 (en) * 2005-09-19 2008-09-11 Basf Aktiengesellschaft Novel Dehydrogenases, the Derivatives thereof, and Method for the Production of Optically Active Alkanols
CN101613672A (en) * 2009-08-04 2009-12-30 南京工业大学 A kind of recombination bacillus coli and construction process thereof of asymmetric conversion preparation (S)-4-chloro-ethyl 3-hydroxybutanoate
CN101993828A (en) * 2010-11-08 2011-03-30 江南大学 Method for improving transformation efficiency of (S)-phenyl glycol by coupling glucose-6-phosphate dehydrogenase and (S)-carbonyl reductase
CN102206686A (en) * 2011-04-19 2011-10-05 华东理工大学 Preparation method of methyl (R)-o-chloromandelate utilizing biocatalytic asymmetric reduction

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LI, LINGLING ET AL.: "Regeneration of catalytic coenzyme of recombinant glucose dehydrogenase", JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY ( NATURE SCIENCE EDITION, vol. 38, no. 3, March 2010 (2010-03-01), pages 112 - 115 AND 132 *

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